Process for producing viral RNA polymerase

Details Process for producing viral RNA polymerase Process for producing viral RNA polymerase

2006-03-21

2006-03-21

Housel, James (Department: 1648)

Chemistry: molecular biology and microbiology

Virus or bacteriophage, except for viral vector or...

Inactivation or attenuation; producing viral subunits

C435S069100, C435S239000, C435S320100

Reexamination Certificate

active

07015025

ABSTRACT:
This invention relates to a method for producing virus RNA polymerases of RNA viruses, more specifically, virus RNA polymerases of RNA viruses free of virus genomic RNA.The methods described in this invention includes the procedures for preparation of cDNAs for the genes for the component proteins of RNA polymerase of an RNA virus, incorporation of the cDNA into baculovirus genome to construct recombinant virus, and the infection of insect cells with the recombinant virus to express RNA polymerase. In this method, it is recommended that all species of the recombinant viruses, each of which is designed for expressing each of the above-mentioned component protein genes of RNA polymerase, are coinfected into insect cells. Thus, cDNA is prepared for each of the component proteins of RNA polymerase and incorporated into baculovirus genome to construct recombinant virus for independently expressing the corresponding protein. In addition, the RNA viruses described above include influenza virus especially. In this method, RNA polymerase may be tagged to facilitate the subsequent purification. Furthermore, the tagged RNA polymerase described above may be purified using an adsorbent to trap the RNA polymerase at the tag. Another subject matter of this invention is to artificially prepare a complex of the component proteins of RNA polymerase of an RNA virus, or, in other words, to prepare RNA polymerase free from RNA virus genomic RNA. The RNA viruses described here include influenza virus, and the component proteins of RNA polymerase include PA, PB1 and PB2.

REFERENCES:
patent: 9-121867 (1997-05-01), None
patent: 11-199489 (1999-07-01), None
Gal-on et al., Arch Virology 2000 vol. 145, pp. 37-50.
Makoto Kobayashi, et al, “Reconstitution of influenza virus RNA polymerase from three subunits expressed using recombinant baculovirus system,” Virus Research, vol. 22, 1992, pp. 235-245.
M. Kobayashi, et al, “Influenza virus PB1 protein is the minimal and essential subunit of RNA polymerase,” Arch. Virol, vol. 141, 1996, pp. 525-539.
Licia Tomei, et al, “Biochemical characterization of a hepatitis C virus RNA-dependent RNA polymerase mutant lacking the C-terminal hydrophobic sequence,” Journal of General Virology, vol. 81, 2000, pp. 759-767.
A. Gal-On, et al, “Characterisation of genetically modified cucumber mosaic virus expressing histidine-tagged 1a and 2a proteins,” Arch. Virol., vol. 145, 2000, pp. 37-50.
Timothy Smith, et al, “Purification and Characterization of the Human Recombinant Histidine-Tagged Prostaglandin Endoperoxide H Synthases-1 and -2,” Archives of Biochemistry and Biophysics, vol. 375, No. 1, 2000, pp. 195-200.
Tetsuya Toyoda, et al, “Molecular assembly of the influenza virus RNA polymerase: determination of the subunit-subunit contact sites,” Journal of General Virology, vol. 77, 1996, pp. 2149-2157.
Ayae Honda, et al, “Purification and Molecular Structure of RNA Polymerase from Influenza Virus A/PR81,” J. Biochem, vol. 107, 1990, pp. 624-628.
Tetsuo Miyamura, “Properties of HCV nonstructural protein,” Annual research report for the 9thyear of the Heisei era. 1997.
Hwang et al., “Expression of Functional Influenza Virus RNA Polymerase in the Methylotrophic YeastPichia pastoris, ” Journal of Virology, 2000, pp. 4074-4084, vol. 74, No. 9, American Society for Microbiology, Washington, D.C.
Honda et al., “Differential Roles of Viral RNA and cRNA in Functional Modulation of the Influenza Virus RNA Polymerase,” Journal of Biological Chemistry, 2001, pp. 31179-31185, vol. 276, No. 33, The American Society for Biochemistry and Molecular Biology, Inc., Baltimore, MD.

Affiliated with

Also associated with

Rating

Process for producing viral RNA polymerase does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Process for producing viral RNA polymerase, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Process for producing viral RNA polymerase will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3549745